In vitro antimicrobial susceptibility of Mycoplasma bovis isolated in Israel from local and imported cattle

Gerchman, Irena; Levisohn, Sharon; Mikula, I.; Lysnyansky, Inna

doi:10.1016/j.vetmic.2009.01.028

Cited by 54 publications

(83 citation statements)

References 25 publications

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“…These included 42 isolates for which the geographic origin, clinical condition, and susceptibility profiles have been described previously (7,10) DNA extraction procedures. Genomic DNA was extracted from different samples, including (i) logarithmic-phase broth culture, (ii) nasal swabs soaked in sterile 1ϫ phosphate-buffered saline (PBS), (iii) transtracheal lavage (TTL) fluid, and (iv) bacterial (mycoplasma and nonmycoplasma) colonies collected from agar plates by swabbing and suspension in sterile PBS.…”

Section: Mycoplasma Bovis and Culture Media A Total Of 133 M Bovis mentioning

confidence: 99%

“…Initially, evaluation of the SNP-real-time PCRs was performed on DNAs extracted from M. bovis strains 8830, 9603, 0962, and 8998 with the known parC genotypes AAT, GAT, AAC, and GAC, and enrofloxacin MIC values of 5, 0.16, 5, and 0.08 g/ml, respectively, as previously reported (7,10). Since the two probes in the same tube are labeled with different dyes (FAM and VIC), the presence of the respective SNP was easily detected from the relative fluorescence intensities of the two fluorophores (Fig.…”

Section: Mycoplasma Bovis and Culture Media A Total Of 133 M Bovis mentioning

confidence: 99%

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Development and Evaluation of a Novel Single-Nucleotide-Polymorphism Real-Time PCR Assay for Rapid Detection of Fluoroquinolone-Resistant Mycoplasma bovis

Shabat¹,

Mikula²,

Gerchman³

et al. 2010

J Clin Microbiol

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Monitoring of the susceptibility of Mycoplasma bovis field isolates to antibiotics is important for the appropriate choice of treatment. However, in vitro susceptibility testing of mycoplasmas is technically demanding and time-consuming, especially for clinical isolates, and is rarely performed in mycoplasma diagnostic laboratories. Thus, the development of methods allowing rapid real-time detection of resistant strains of M. bovis in clinical samples is a high priority for successful treatment. In this study, a novel TaqMan singlenucleotide-polymorphism (SNP) real-time PCR assay, which enables the rapid identification of M. bovis strains with different susceptibilities to fluoroquinolones, was developed and evaluated. The TaqMan SNP real-time PCR assay is based on the amplification of a 97-bp fragment of the parC quinolone resistance-determining region (QRDR) and allows the specific detection of four possible genotypes: GAC or GAT (susceptible to fluoroquinolones) and AAC or AAT (resistant to fluoroquinolones). Four TaqMan minor groove binder (MGB) probes identifying 1-base mismatches were designed and applied in a dual-probe assay with two reaction tubes. The TaqMan SNP real-time PCRs developed are highly specific for M. bovis, with a detection limit of 5 fg/l (about 5 M. bovis genomes). In addition, all four SNP real-time PCR tests have almost the same efficiency (97.7% [GAC], 94% [AAC], 99.99% [GAT], and 98% [AAT]). Taken together, the data suggest that this SNP real-time PCR assay has potential as a routine diagnostic test for the detection of decreased susceptibility of M. bovis to fluoroquinolones.Mycoplasma bovis is an important and emerging cause of bovine respiratory disease (BRD), calf pneumonia, mastitis, arthritis, and otitis media, as well as various less common presentations (11). Clinical disease associated with M. bovis is often chronic, debilitating, and poorly responsive to antimicrobial therapy, resulting in significant economic loss, the full extent of which is sometimes difficult to estimate (5, 11). Moreover, since there is no effective vaccine for M. bovis, use of antibiotics is the main control method for this infection.Fluoroquinolones are broad-spectrum antimicrobials highly effective for the treatment of a variety of clinical and veterinary infections and frequently used for the treatment of M. bovis infection in cattle. Their antibacterial activity is due mainly to inhibition of DNA replication. A major mechanism of fluoroquinolone resistance in prokaryotes (including Mollicutes) involves alterations in the target enzymes DNA gyrase and DNA topoisomerase IV, thereby altering affinity for the drug (2, 8). The mutations associated with resistance have been localized within the quinolone resistance-determining regions (QRDRs) of DNA gyrase subunits GyrA and GyrB and/or topoisomerase IV subunits ParC and ParE.Recently, the QRDRs of these genes in 42 M. bovis clinical isolates exhibiting various levels of susceptibility to fluoroquinolones were characterized (10). The data showed that 10/...

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Section: Mycoplasma Bovis and Culture Media A Total Of 133 M Bovis mentioning

confidence: 99%

Section: Mycoplasma Bovis and Culture Media A Total Of 133 M Bovis mentioning

confidence: 99%

Development and Evaluation of a Novel Single-Nucleotide-Polymorphism Real-Time PCR Assay for Rapid Detection of Fluoroquinolone-Resistant Mycoplasma bovis

Shabat¹,

Mikula²,

Gerchman³

et al. 2010

J Clin Microbiol

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“…Infrequent or no resistance has been reported to enrofloxacin or florfenicol (Francoz et al, 2005, Gerchman et al, 2009, Hirose et al, 2003, Rosenbusch et al, 2005, Siugzdaite et al, 2012. Successful treatment of M. bovis pneumonia and arthritis in calves using valnemulin in the milk from four days of age for three weeks has also been reported (Stipkovits et al, 2001).…”

Section: Disease Prevention and Controlmentioning

confidence: 99%

“…In fact Gerchman et al (2009) suggests that M. bovis is the most common bacterium identified in feedlot cattle affected by chronic pneumonia and in veal calves with fatal bronchopneumonia. However the causative nature, of M. bovis in cattle older than 6 months of age is still not well understood.…”

Section: Mycoplasma Bovis Associated Pneumoniamentioning

confidence: 99%

“…In addition, in the past decade, in vitro susceptibility studies have shown an increased resistance to antimicrobials commonly used to treat infections associated with M. bovis; in particular oxytetracyclines, spectinomycin, tilmicosin and chlortetracycline (Francoz et al, 2005, Gerchman et al, 2009, Hirose et al, 2003, Rosenbusch et al, 2005, Siugzdaite et al, 2012. Infrequent or no resistance has been reported to enrofloxacin or florfenicol (Francoz et al, 2005, Gerchman et al, 2009, Hirose et al, 2003, Rosenbusch et al, 2005, Siugzdaite et al, 2012.…”

Section: Disease Prevention and Controlmentioning

confidence: 99%

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Mycoplasma bovis in Australian feeder cattle

Schibrowski¹

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Mycoplasma bovis can be a commensal bacterial inhabitant of the upper respiratory tract of healthy bovines. However, under certain circumstances and conditions that are incompletely understood, M. bovis can be associated with a number of clinical syndromes, one of which is bovine respiratory disease (BRD). In Australia, there is little information on the prevalence and epidemiology M. bovis in beef production systems. The overall aim of this thesis was to help close this knowledge gap.Commercially available serological diagnostic methods for the detection of M. bovis-specific antibody have not been validated in the Australian cattle population. Arguably, more importantly, however is the fact that diagnostic sensitivity (Se) and specificity (Sp) and / or analytical sensitivity (ASe) and specificity (ASp) estimates for commercially available M. bovis sero-diagnostic tests are not available. 28% (95% CI: 1% -92%). However, in terms of Sp the commercial ELISAs out performed western blotting; BioK302: 96% (95% CI: 87% -99%); BioK260:100% (95% CI: 93% -100%) and western blotting: 88% (95% CI: 56% -98%) respectively. In terms of both Se and Sp combined, none of the methods assessed here performed well. As such, when interpreting the results obtained from these ELISAs, the potential impact imperfect Se and / or Sp can have needs to be considered.Despite the poor performance of the BioK302 and BioK260 ELISAs particularly in terms of Se, these tests were used to determine the sero-prevalence of M. bovis antibody in Australian feeder cattle at feedlot induction and draft (approximately 42 days on feed). Paired bovine sera from 1,354 randomly selected animals were sourced from a bovine serum bank (n = 35,160) collected as part of another study, The National Bovine Respiratory Disease Initiative. As the classification of the result of a single serum sample could differ between the BioK302 and BioK260, sero-prevalence estimates were obtained using the individual ELISA results in addition to series and parallel interpretation of the two ELISA kit results. The apparent sero-prevalence of M. bovis-specific antibody at feedlot induction estimates ranged from 2.4% -6.8% (95% CI: 1.3% -8.6%) and the apparent sero-prevalence estimates at draft ranged from 22.4% -44.2% (95% CI: 19.1% -48.6%). The level of sero-conversion to M. bovis between induction and draft was also determined. Only animals that were classified as sero-negative at induction were included in sero-conversion analyses. In the current study, 24.3% (95% iii CI: 20.8% -27.7%) and 19.5% (95% CI: 17.3% -21.7%) of animals sero-converted to M. bovis between induction and draft based on the BioK302 and dichotomised BioK260 ELISA results respectively.Using these ELISA results, an assessment of putative risk factors associated with sero-positivity at feedlot induction or sero-conversion between induction and draft was performed. An increased risk of sero-positivity at feedlot induction was found to be associated with; (i) history of exposure to a saleyard at least 27 d...

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Diseases of the Respiratory System

2017

Veterinary Medicine

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In vitro antimicrobial susceptibility of Mycoplasma bovis isolated in Israel from local and imported cattle

Cited by 54 publications

References 25 publications

Development and Evaluation of a Novel Single-Nucleotide-Polymorphism Real-Time PCR Assay for Rapid Detection of Fluoroquinolone-Resistant Mycoplasma bovis

Development and Evaluation of a Novel Single-Nucleotide-Polymorphism Real-Time PCR Assay for Rapid Detection of Fluoroquinolone-Resistant Mycoplasma bovis

Mycoplasma bovis in Australian feeder cattle

Diseases of the Respiratory System

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